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Yosuke Makuuchi,
Kazufumi Honda,
Yoshiaki Osaka,
Ken Kato,
Takashi Kojima,
Hiroyuki Daiko,
Hiroyasu Igaki,
Yoshinori Ito,
Sumito Hoshino,
Shingo Tachibana,
Takafumi Watanabe, Koh Furuta,
Shigeki Sekine,
Tomoko Umaki,
Yukio Watabe,
Nami Miura,
Masaya Ono,
Akihiko Tsuchida,
Tesshi Yamada
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ABSTRACT: Preoperative chemoradiotherapy has been shown to improve the outcome of patients with esophageal cancer, but because response to this therapy varies, it is desirable to identify in advance individuals who would be unlikely to benefit in order to avoid unnecessary adverse drug effects. The serum profiles of 84 cytokines and related proteins were determined in 37 patients with esophageal squamous cell carcinoma who received identical neoadjuvant preoperative chemoradiotherapy regimens and underwent surgical resection. Histological response to this therapy was assessed in surgically resected specimens. The serum soluble interleukin-6 receptor level was significantly higher in 30 patients who failed to achieve a histological complete response (P = 0.005). Multivariate analysis revealed that the increased level of soluble interleukin-6 receptor was one of several significant independent predictors of an unfavorable outcome (hazard ratio, 2.87; P = 0.017). The increased level of this cytokine in patients who did not obtain a complete response was reproducibly observed in an independent cohort of 34 patients. Esophageal squamous cell carcinoma patients with an increased serum level of soluble interleukin-6 receptor are predicted to respond poorly to preoperative chemoradiotherapy, and therefore their exclusion from this treatment may be considered. Persistent systemic inflammation is implicated as a possible mechanism of resistance to this therapy. This article is protected by copyright. All rights reserved.
Cancer Science 05/2013; · 3.33 Impact Factor
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ABSTRACT: Direct sequencing (DS) has often been used for detection of v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations. High-resolution melting analysis (HRMA) is another method to detect mutations by using a light scanner, and is more rapid, lower in cost, and more sensitive than DS. We confirmed correspondence between DS and HRMA for KRAS mutation detection in colorectal cancer (CRC).
From 102 patients with CRC, intended to receive cetuximab treatment at the National Cancer Center Hospital between September 2008 and July 2009, formalin-fixed paraffin-embedded tissues were retrospectively analyzed for KRAS status using HRMA and DS. Cetuximab efficacy was evaluated.
Success rates of analysis by DS and HRMA were 100 out of the 102 patients (98.0%) and 99 out of the 102 patients (97.1%), respectively. The cases which failed by one method were analyzed by the other. KRAS mutant-type was detected by DS in 47 out of 100 samples (47.0%), and by HRMA in 45 out of 99 samples (45.5%). The concordance between the two methods was 94.8%. Forty-six out of the 53 patients with wild-type KRAS, as detected by DS received cetuximab and the response and disease control rates were 19.6% and 63.0%, respectively. With a median follow-up of 8.8 months, the median progression-free survival was 5.6 months and median overall survival was 11.1 months. The efficacy of two discordant cases which received cetuximab showed that the best responses were stable disease and progressive disease in one each, progression-free survival was 2.9 and 1.1 months and overall survival was 5.3 and 1.2 months, respectively.
HRMA is a useful optional method for detection of KRAS mutations in CRC in light of accuracy, cost performance and rapidity.
Anticancer research 05/2013; 33(5):2129-34. · 1.73 Impact Factor
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ABSTRACT: AIMS: In lung cancer with anaplastic lymphoma kinase (ALK) gene rearrangement, accurate diagnosis is essential to identify patients who can be treated with a specific kinase inhibitor. Sensitive ALK immunostaining can provide nearly complete concordance with gene rearrangement status and is expected to serve as a surrogate biomarker for tailored treatment with an ALK inhibitor. In the present report, we describe aberrant ALK expression in a small proportion of pulmonary neuroendocrine carcinomas (NECs) that do not have ALK gene alteration. METHODS: A total of 227 pulmonary NECs were assembled on tissue microarrays and were subjected to highly sensitive ALK staining methods. RESULTS: We observed focal positivity with heterogeneous intensity in 2 (2.9%) of 69 small-cell carcinomas and 1 (0.9%) of 106 large-cell NECs. In contrast, 52 carcinoid tumours were all negative for ALK expression. Neither ALK rearrangement nor amplification was observed using fluorescence in situ hybridisation, and no somatic mutation was detected in three ALK positive NECs. CONCLUSIONS: Thus, this aberrant expression is probably of a wild-type ALK and a potential pitfall when implementing sensitive ALK immunohistochemistry in the molecular diagnosis of lung cancer.
Journal of clinical pathology 04/2013; · 2.43 Impact Factor
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Shigeki Morita,
Akihiko Yoshida,
Akiteru Goto,
Satoshi Ota,
Koji Tsuta,
Karin Yokozawa,
Hisao Asamura,
Jun Nakajima,
Daiya Takai,
Masaya Mori,
Teruaki Oka,
Junichi Tamaru,
Shinji Itoyama, Koh Furuta,
Masashi Fukayama,
Hitoshi Tsuda
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ABSTRACT: Low-grade lung adenocarcinoma of fetal lung type, which is well characterized by its unique clinicopathologic and molecular features, is recognized as a distinct variant of lung cancer. In contrast, high-grade lung adenocarcinoma with fetal lung-like morphology (HG-LAFM) has not been studied widely. To characterize this subset better, we analyzed 17 high-grade adenocarcinomas with at least focal component resembling a developing epithelium in the pseudoglandular phase of the fetal lung. These rare (ca. 0.4%) carcinomas occurred predominantly in elderly men with a heavy smoking history, who showed elevated serum α-fetoprotein in 4 of 5 cases tested. Histologic examination revealed a fetal lung-like component as a focal finding accounting for 5% to 60% of the total tumor volume. It was invariably admixed with tissues having a morphology not resembling that of a fetal lung. A coexisting non-fetal lung-like element was quite heterogenous in appearance, showing various growth patterns. However, clear-cell (88%), hepatoid (29%), and large cell neuroendocrine carcinoma (24%) histology seemed overrepresented. HG-LAFM was characterized immunohistochemically by frequent expression of α-fetoprotein (41%), glypican-3 (88%), SALL-4 (59%), neuroendocrine markers (82%), CDX-2 (35%), and p53 (65%). HG-LAFM was molecularly heterogenous in that EGFR or KRAS mutation was observed in 22% of cases tested for both. Our data indicate that HG-LAFMs might form a coherent subgroup of lung adenocarcinomas. However, the uniformly focal nature of the fetal lung-like element, widely diverse coexisting non-fetal lung-like histology, and inhomogenous molecular profiles lead us to believe that HG-LAFM is best regarded as a morphologic pattern showing characteristic association with several clinicopathologic parameters rather than a specific tumor entity.
The American journal of surgical pathology 04/2013; · 4.06 Impact Factor
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Takafumi Suzuki,
Tatsuhiro Shibata,
Kai Takaya,
Kouya Shiraishi,
Takashi Kohno,
Hideo Kunitoh,
Koji Tsuta, Koh Furuta,
Koichi Goto,
Fumie Hosoda,
Hiromi Sakamoto,
Hozumi Motohashi,
Masayuki Yamamoto
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ABSTRACT: Transcription factor Nrf2 (NF-E2-related factor-2) is essential for oxidative and electrophilic stress responses. While it has been well characterized that Nrf2 activity is tightly regulated at the protein level through proteasomal degradation via Keap1 (Kelch-like ECH-associated protein 1)-mediated ubiquitination, not much attention has been paid to the supply-side of Nrf2, especially regulation of Nrf2 gene transcription. Using genetically engineered mouse models, here we report that manipulation of Nrf2 transcription is effective in changing the final Nrf2 protein level and activity of cellular defense against oxidative stress even in the presence of Keap1 and under efficient Nrf2 degradation. In excellent agreement, we found that minor A/A homozygotes of a single nucleotide polymorphism (SNP) in human NRF2 upstream-promoter region (rs6721961) exhibited significantly diminished NRF2 gene expression and consequently an increased risk of lung cancer, especially in ever-smokers. Our results support the notion that in addition to control over proteasomal degradation and derepression from the degradation/repression, the transcriptional level of the Nrf2 gene acts as another important regulatory point to define cellular Nrf2 levels. These results thus verify the critical importance of human SNPs that influence transcription levels of the NRF2 gene for future personalized medicine.
Molecular and cellular biology 04/2013; · 6.06 Impact Factor
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Nobuhiro Hiramoto,
Yukio Kobayashi,
Junko Nomoto,
Dai Maruyama,
Takashi Watanabe,
Naobumi Tochigi, Koh Furuta,
Ken Takeda,
Hirokazu Chuman,
Shigeki Yagyu,
Hajime Hosoi,
Kensei Tobinai
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ABSTRACT: We report the case of a patient in whom the diagnosis of Ewing sarcoma arising from a soft tissue was made after successful treatment of diffuse large B-cell lymphoma. A 65-year-old woman presented with a rapidly growing mass in her left scapular region 8 years after successful chemotherapy with the cyclophosphamide, hydroxydaunomycin hydrochloride, vincristine, prednisolone regimen for diffuse large B-cell lymphoma. Computed tomographic examination and magnetic resonance imaging of the thorax revealed an intramuscular tumour measuring 40 mm in size in the left scapular region. Histopathological examination of an open biopsy specimen revealed a small round cell tumour that showed positive staining for CD99. Fluorescence in situ hybridization showed a split signal by a break-apart probe for the EWS gene in chromosome 22q12. Reverse transcriptase-polymerase chain reaction confirmed the expression of EWS-FLI1 fusion transcripts. Based on these findings, the patient was diagnosed as having secondary Ewing sarcoma. Despite adjuvant chemotherapy, however, she died of pulmonary metastases 2 years after the diagnosis of Ewing sarcoma. Therapy-related haematological malignancies with balanced translocations have been reported previously. A mechanism similar to that underlying the development of secondary malignancy might explain the occurrence of this solid cancer.
Japanese Journal of Clinical Oncology 03/2013; · 1.78 Impact Factor
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ABSTRACT: Recent discovery of ROS1 gene fusion in a subset of lung cancers has raised clinical interest, because ROS1 fusion-positive cancers are reportedly sensitive to kinase inhibitors. To better understand these tumors, we examined 799 surgically resected non-small cell lung cancers by reverse transcriptase polymerase chain reaction and identified 15 tumors harboring ROS1 fusion transcripts (2.5% of adenocarcinomas). The most frequent fusion partner was CD74 followed by EZR. The affected patients were often younger nonsmoking female individuals, and they had overall survival rates similar to those of the ROS1 fusion-negative cancer patients. All the ROS1 fusion-positive tumors were adenocarcinomas except 1, which was an adenosquamous carcinoma. Histologic examination identified an at least focal presence of either solid growth with signet-ring cells or cribriform architecture with abundant extracellular mucus in 53% of the cases. These 2 patterns are reportedly also characteristic of anaplastic lymphoma kinase (ALK)-rearranged lung cancers, and our data suggest a phenotypic resemblance between the ROS1-rearranged and ALK-rearranged tumors. All tumors except 1 were immunoreactive to thyroid transcription factor-1. Fluorescence in situ hybridization using ROS1 break-apart probes revealed positive rearrangement signals in 23% to 93% of the tumor cells in ROS1 fusion-positive cancers, which were readily distinguished using a 15% cutoff value from 50 ROS1 fusion-negative tumors tested, which showed 0% to 6% rearrangement signals. However, this perfect test performance was achieved only when isolated 3' signals were included along with classic split signals in the definition of rearrangement positivity. Fluorescence in situ hybridization signal patterns were unrelated to 5' fusion partner genes. All ROS1 fusion-positive tumors lacked alteration of EGFR, KRAS, HER2, ALK, and RET genes.
The American journal of surgical pathology 02/2013; · 4.06 Impact Factor
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ABSTRACT: Rhabdomyosarcoma is a rare soft tissue sarcoma that typically affects children, adolescents, and young adults. Despite treatment via a multidisciplinary approach, the prognosis of advance-stage rhabdomyosarcomas remains poor, and a new treatment strategy is needed. Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase that is a potential target for specific inhibitors. In this study, we investigated 116 rhabdomyosarcomas using a polymer-based ALK immunostaining method and correlated the results with clinicopathological parameters. In addition, we examined ALK status using dual-color fluorescence in situ hybridization, PCR, and sequencing. In immunohistochemical analysis, ALK was detected in 2 (6%) of 33 embryonal rhabdomyosarcomas, 42 (69%) of 61 alveolar rhabdomyosarcomas, and 0 (0%) of 22 other subtypes, including pleomorphic, adult-spindle-cell/sclerosing, and epithelioid variants. Compared with ALK-negative alveolar rhabdomyosarcomas, ALK-positive ones are presented with metastatic spread more frequently and showed a greater extent of myogenin reactivity. Overall survival was not associated with ALK expression. FOXO1 rearrangement was significantly associated with ALK immunoreactivity. The median ALK copy number was greater in ALK-positive tumors than in ALK-negative tumors. Most (93%) cases tested showed no selective increase in the ALK gene dosage. ALK selective amplification and low-level selective gain were noted in one and three cases, respectively. Further, a high-polysomy pattern (≥4 ALK copies in ≥40% of cells) was observed in seven cases. A significant increase in the ALK copy number was exclusive to the ALK-immunopositive cohort, but it was uncommon, accounting for only 30% of the 37 ALK-positive rhabdomyosarcomas. ALK gene rearrangement was not observed in either cohort, while an ALK somatic mutation (I1277T) was found in one ALK-negative embryonal case. Although it remains controversial whether ALK expression without gene rearrangement is therapeutically relevant, this comprehensive analysis may help future studies on the utility of ALK-targeted therapy for patients with rhabdomyosarcoma.Modern Pathology advance online publication, 11 January 2013; doi:10.1038/modpathol.2012.222.
Modern Pathology 01/2013; · 4.79 Impact Factor
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ABSTRACT: Insulin-like growth factor-1 receptor (IGF-1R) is a tyrosine kinase receptor implicated in the pathogenesis of several malignancies and is potentially an attractive target for anticancer treatment. In this study, we included 379 patients who underwent surgical resection (179 diagnosed as having adenocarcinoma [ADC]; 150, squamous cell carcinoma [SCC]; 41, sarcomatoid carcinoma and 9, large cell carcinoma). IGF-1R expression and gene copy number were assessed by immunohistochemistry and bright-field in situ hybridization (BISH), respectively. IGF-1R expression in non-small cell lung carcinoma was observed in 41.4% of samples and was more prevalent in SCC (69.3%) than in ADC (25.1%), large cell carcinoma (33.3%), and sarcomatoid carcinoma (12.2%) (P < .001). Among ADCs, most mucinous ADCs (75%) showed strong membranous staining with the IGF-1R antibody. Compared with protein expression, IGF-1R gene alteration was rare (8.4%). A statistically significant correlation between IGF-1R expression and positive IGF-1R BISH was observed (γ = 0.762, P < .001). IGF-1R-positive tumors were more common in smokers (P = .004), and these tumors were larger (P = .006) than the IGF-1R-negative tumors. IGF-1R BISH positivity was not correlated with any clinicopathologic factor. IGF-1R expression and IGF-1R BISH positivity were not correlated with overall survival. IGF-1R is highly expressed in SCC and mucinous ADC, although copy number alterations in the IGF-1R gene were rare. These findings may have important implications for future anti-IGF-1R therapeutic approaches.
Human pathology 12/2012; · 3.03 Impact Factor
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Kohei Tada,
Sung-Won Kim,
Yoshitaka Asakura,
Nobuhiro Hiramoto,
Kimikazu Yakushijin,
Saiko Kurosawa,
Kinuko Tajima,
Shin-ichiro Mori,
Yuji Heike,
Ryuji Tanosaki,
Akiko Miyagi Maeshima,
Hirokazu Taniguchi, Koh Furuta,
Yoshikazu Kagami,
Yoshihiro Matsuno,
Kensei Tobinai,
Yoichi Takaue,
Takahiro Fukuda
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ABSTRACT: The outcome after allogeneic hematopoietic stem cell transplantation (allo-HCT) for diffuse large B-cell lymphoma (DLBCL) associated with follicular lymphoma (FL), which includes DLBCL with pre- or co-existing FL, remains controversial, and few previous reports have compared the outcomes after allo-HCT for FL, DLBCL associated with FL, and de novo DLBCL. We retrospectively analyzed 97 consecutive patients with FL (n = 46), DLBCL associated with FL (n = 22), or de novo DLBCL (n = 29) who received allo-HCT at our institute between 2000 and 2010. With a median follow-up of 53 months, the 5-year overall survival (OS) and progression-free survival (PFS) were, respectively, 77% and 70% for FL, 62% and 57% for DLBCL associated with FL, and 26% and 23% for de novo DLBCL. The 5-year cumulative incidences of non-relapse mortality and disease progression/relapse were, respectively, 16% and 15% for FL, 19% and 24% for DLBCL associated with FL, and 36% and 41% for de novo DLBCL. By a multivariate analysis, the OS and PFS for DLBCL associated with FL were significantly better than those for de novo DLBCL, whereas they were not significantly different from those for FL. These results suggest that allo-HCT may be a promising option for patients with not only advanced FL but also DLBCL associated with FL.
American Journal of Hematology 04/2012; 87(8):770-5. · 4.67 Impact Factor
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ABSTRACT: Ewing sarcoma is a high-grade round cell sarcoma that affects bones and soft tissues in children and young adults. Its diagnosis can be challenging, and the differential diagnoses include a wide variety of small round cell tumors. CD99 and FLI-1 are the currently accepted immunohistochemical markers for Ewing sarcoma, but their accuracy has been controversial. NKX2.2 is a homeodomain-containing transcription factor that plays a critical role in neuroendocrine/glial differentiation. The NKX2.2 gene was recently identified as a target of EWS-FLI-1, the fusion protein specific to Ewing sarcoma, and was shown to be differentially upregulated in Ewing sarcoma on the basis of array-based gene expression analysis. However, the immunohistochemical diagnostic potential of this marker has not been tested. We immunostained representative sections of 30 genetically confirmed Ewing sarcomas and 130 non-Ewing small round cell tumors by using an antibody to NKX2.2. Nuclear staining in at least 5% of the cells was deemed positive. Twenty-eight (93%) of the 30 Ewing sarcomas were positive for NKX2.2. The staining was diffuse (>50%) in all the positive cases and was moderate or strong in intensity for most cases (25 of 28). NKX2.2 was also positive in 14 non-Ewing tumors, including all the olfactory neuroblastomas and a minor subset of small cell carcinomas, synovial sarcomas, mesenchymal chondrosarcomas, and malignant melanomas. All the other non-Ewing tumors tested were negative for this marker. NKX2.2 is a valuable marker for Ewing sarcoma, with a sensitivity of 93% and a specificity of 89%, and aids in the differential diagnosis of small round cell tumors.
The American journal of surgical pathology 03/2012; 36(7):993-9. · 4.06 Impact Factor
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Takashi Kohno,
Hitoshi Ichikawa,
Yasushi Totoki,
Kazuki Yasuda,
Masaki Hiramoto,
Takao Nammo,
Hiromi Sakamoto,
Koji Tsuta, Koh Furuta,
Yoko Shimada, [......],
Itaru Yamanaka,
Yasuhito Arai,
Shun-Ichi Watanabe,
Ikuo Sekine,
Seishi Ogawa,
Curtis C Harris,
Hitoshi Tsuda,
Teruhiko Yoshida,
Jun Yokota,
Tatsuhiro Shibata
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ABSTRACT: We identified in-frame fusion transcripts of KIF5B (the kinesin family 5B gene) and the RET oncogene, which are present in 1-2% of lung adenocarcinomas (LADCs) from people from Japan and the United States, using whole-transcriptome sequencing. The KIF5B-RET fusion leads to aberrant activation of RET kinase and is considered to be a new driver mutation of LADC because it segregates from mutations or fusions in EGFR, KRAS, HER2 and ALK, and a RET tyrosine kinase inhibitor, vandetanib, suppresses the fusion-induced anchorage-independent growth activity of NIH3T3 cells.
Nature medicine 01/2012; 18(3):375-7. · 27.14 Impact Factor
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ABSTRACT: Development of tailored treatment based on immunohistochemical profiles (IPs) of tumors for cancers of unknown primary is needed.
We developed an algorithm based on primary known adenocarcinoma for testing sensitivity and specificity. Formalin-fixed paraffin-embedded tissue samples from 71 patients of unfavorable subsets of unknown primary adenocarcinoma were obtained. We examined 15 molecular markers using the algorithm incorporating these IPs and classified the tumours into 9 subsets based on the primary tumour site. The sensitivity and specificity of this algorithm were 80.3% and 97.6%, respectively. Apparent primary sites were lung in 17 patients, digestive organs in 13, gynecological organs in 9, prostate in 7, liver or kidney in 6, breast in 4, urothelial organ in 2, biliary tract and pancreatic profile in none, and unclassified in 13. The response rate to chemotherapy was highest for the gynecological IPs. Patients with gynecological or lung cancer IPs had longer median progression-free survival than those with others: 11.2 months for gynecological IPs (p<0.001) and 6.8 months for lung IPs (p = 0.05). Lung, digestive, prostate, and gynecological profiles were associated with significantly longer median survival time than the other profiles. Multivariate analysis confirmed that the IPs were independent prognostic factors for survival.
The IPs identified in this study can be used to further stratify patient prognosis for unfavorable subsets of unknown primary adenocarcinoma.
PLoS ONE 01/2012; 7(1):e31181. · 4.09 Impact Factor
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ABSTRACT: The hepatocyte growth factor/MET pathway has been shown to cause tumor progression in several types of carcinomas. The aim of this study was to examine the correlations between c-MET/phospho-MET expression as well as MET gene copy number alterations and overall survival (OS) in non-small cell lung carcinomas (NSCLCs).
We analyzed 906 NSCLCs including 704 adenocarcinomas (ADCs), 150 squamous cell carcinomas (SCCs), 43 sarcomatoid carcinomas, and 9 large cell carcinomas. The mutational status of epidermal growth factor receptor and K-ras and anaplastic lymphoma kinase rearrangements were retrospectively examined. We performed immunohistochemistry to detect c-MET/phospho-MET expression and MET gene copy number using bright-field in situ hybridization (BISH).
c-MET/phospho-MET expression and MET BISH positivity were observed in 22.2%, 5.6%, and 10.9% of NSCLCs, respectively; they were more prevalent in ADCs (27.3%, 6.9%, and 11.5%, respectively) and sarcomatoid carcinomas (20.9%, 9.3%, and 36.6%, respectively) than in SCCs and large cell carcinomas. Among ADCs, poorly differentiated cases exhibited c-MET expression and MET BISH positivity more commonly than well-differentiated ones. An analysis of all patients revealed that c-MET/phospho-MET expression and MET BISH positivity were not correlated with OS. However, when SCC cases were excluded, both univariate (p=0.019) and multivariate (p=0.020) analyses revealed a significant correlation between MET BISH positivity and OS.
c-MET/phospho-MET expression and MET BISH positivity differed according to histological type. Among ADCs, c-MET expression and MET BISH positivity were more common in poorly differentiated cases. MET BISH positivity was an independent prognostic factor in nonsquamous NSCLCs.
Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 12/2011; 7(2):331-9. · 4.55 Impact Factor
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Hirokazu Okayama,
Takashi Kohno,
Yuko Ishii,
Yoko Shimada,
Kouya Shiraishi,
Reika Iwakawa, Koh Furuta,
Koji Tsuta,
Tatsuhiro Shibata,
Seiichiro Yamamoto, [......],
Hiromi Sakamoto,
Kensuke Kumamoto,
Seiichi Takenoshita,
Noriko Gotoh,
Hideaki Mizuno,
Akinori Sarai,
Shuichi Kawano,
Rui Yamaguchi,
Satoru Miyano,
Jun Yokota
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ABSTRACT: Activation of the EGFR, KRAS, and ALK oncogenes defines 3 different pathways of molecular pathogenesis in lung adenocarcinoma. However, many tumors lack activation of any pathway (triple-negative lung adenocarcinomas) posing a challenge for prognosis and treatment. Here, we report an extensive genome-wide expression profiling of 226 primary human stage I-II lung adenocarcinomas that elucidates molecular characteristics of tumors that harbor ALK mutations or that lack EGFR, KRAS, and ALK mutations, that is, triple-negative adenocarcinomas. One hundred and seventy-four genes were selected as being upregulated specifically in 79 lung adenocarcinomas without EGFR and KRAS mutations. Unsupervised clustering using a 174-gene signature, including ALK itself, classified these 2 groups of tumors into ALK-positive cases and 2 distinct groups of triple-negative cases (groups A and B). Notably, group A triple-negative cases had a worse prognosis for relapse and death, compared with cases with EGFR, KRAS, or ALK mutations or group B triple-negative cases. In ALK-positive tumors, 30 genes, including ALK and GRIN2A, were commonly overexpressed, whereas in group A triple-negative cases, 9 genes were commonly overexpressed, including a candidate diagnostic/therapeutic target DEPDC1, that were determined to be critical for predicting a worse prognosis. Our findings are important because they provide a molecular basis of ALK-positive lung adenocarcinomas and triple-negative lung adenocarcinomas and further stratify more or less aggressive subgroups of triple-negative lung ADC, possibly helping identify patients who may gain the most benefit from adjuvant chemotherapy after surgical resection.
Cancer Research 11/2011; 72(1):100-11. · 7.86 Impact Factor
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ABSTRACT: The purpose of this study was to contribute to the quality management in the fields of clinical studies and clinical trials. To accomplish this purpose we tried to make guidelines for nationwide standardization of clinical tests. This study was performed by multidisciplinary personnel such as medical doctors, medical technologist, CRC, and representatives of pharmaceutical companies and/or clinical laboratory testing companies. The study was also focused on two fields of clinical tests such as pre-analytical and post-analytical phase. Each member shared various incidents and tried to make solutions. In post-analytical area in particular, further discussion concerning CDISC and SS-MIX was done. Based on these efforts, a tentative guideline was made.
Rinsho byori. The Japanese journal of clinical pathology 10/2011; 59(10):978-87.
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Akihiko Yoshida,
Koji Tsuta,
Hiroaki Nitta,
Yutaka Hatanaka,
Hisao Asamura,
Ikuo Sekine,
Thomas M Grogan,
Masashi Fukayama,
Tatsuhiro Shibata, Koh Furuta,
Takashi Kohno,
Hitoshi Tsuda
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ABSTRACT: A subset of lung cancers harbors an EML4-ALK (echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase) gene fusion, and detecting this subset may hold therapeutic implications. Many prior studies used fluorescence in situ hybridization (FISH) analysis for this detection, but FISH may have disadvantages including signal decay and dark-field examination that may obscure tissue architecture. In this study, we explored the potential of the ALK-break-apart chromogenic in situ hybridization (CISH) method to detect ALK-rearranged lung cancer.
We examined 15 lung adenocarcinomas with reverse-transcriptase polymerase chain reaction-proven EML4-ALK fusion transcripts and 30 ALK-negative cases. One hundred tumor cells were evaluated by CISH and FISH for each case, and a detailed signal profile was recorded and compared.
CISH preserved tissue architecture and cytomorphology considerably and facilitated the signal evaluation using a routine light microscope. Positive rearrangement signals (splits or isolated 3' signals) were identified in 13 to 78% (mean ± SD, 41% ± 19%) of tumor cells in the ALK-positive cohort and in 0 to 15% (mean ± SD, 6% ± 4%) of cells in the ALK-negative cohort. The two groups were best separated by a cutoff value of 20%, with a sensitivity of 93% and a specificity of 100%. The only false-negative tumor having only 13% CISH-positive cells displayed predominantly (76%) isolated 5' signals unaccompanied by 3' signals. FISH showed largely similar signal profiles, and the results were completely concordant with CISH.
We have successfully introduced CISH for diagnosing EML4-ALK-positive lung adenocarcinoma. This method allows simultaneous visualization of genetics and tumor cytomorphology and facilitates the molecular evaluation and could be applicable in clinical practice to detect lung cancer that may be responsive to ALK inhibitors.
Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 10/2011; 6(10):1677-86. · 4.55 Impact Factor
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Akihiko Yoshida,
Koji Tsuta,
Harumi Nakamura,
Takashi Kohno,
Fumiaki Takahashi,
Hisao Asamura,
Ikuo Sekine,
Masashi Fukayama,
Tatsuhiro Shibata, Koh Furuta,
Hitoshi Tsuda
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ABSTRACT: A subset (1% to 5%) of non-small-cell lung carcinomas harbors the EML4-ALK fusion gene. Data from previous studies on the histomorphology of ALK-rearranged lung cancer are inconsistent, and the specific histologic parameters that characterize this subset and how accurately such parameters predict underlying ALK abnormality remain uncertain. To answer these questions, we performed a comprehensive histologic analysis of 54 surgically resected, extensively sampled ALK-rearranged lung carcinomas and compared them with 100 consecutive resections of ALK-wild-type lung cancers. All 54 cases showed at least a focal adenocarcinoma component, and 3 and 2 cases had additional squamous and sarcomatoid differentiation, respectively. Solid or acinar growth pattern, cribriform structure, presence of mucous cells (signet-ring cells or goblet cells), abundant extracellular mucus, lack of lepidic growth, and lack of significant nuclear pleomorphism were more common in ALK-positive cancers. Two recognizable constellations of findings, a solid signet-ring cell pattern and a mucinous cribriform pattern, were present at least focally in the majority (78%) of ALK-positive tumors, but were rare (1%) in ALK-negative tumors. Multivariate analysis showed that a combination of these 2 patterns was the most powerful histologic indicator of ALK rearrangement. Characteristic histologies were present both in primary sites and in metastases. Thus, histologic findings may help to identify cases for ALK testing. However, none of the histologic parameters were completely sensitive or specific to ALK rearrangement, and histomorphology should not replace confirmatory molecular or immunohistochemical studies. ALK-positive cancers commonly showed coexpression of thyroid transcription factor-1 and p63, and its significance is currently unclear.
The American journal of surgical pathology 08/2011; 35(8):1226-34. · 4.06 Impact Factor
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ABSTRACT: We established our bio-repository in October 2002. One of the unique aspects of our bio-repository is that it is based on post-clinical test samples. Although the post-clinical test sample-based storage is beneficial because ample clinical information is available for each sample and samples are to be refreshed regularly, many bio-bankers are hesitant to store them because of the possible problems of de-identification procedures and sample quality. Currently, we have two different types of sample, not de-identified status and de-identified status. Most of the samples for storage are not de-identified status. A portion of the samples are transferred to new tubes before and after being frozen, without clinical patient identifications and the status is de-identified. This tube transfer is the only de-identification procedure in our bio-repository so far. After a procedure of de-identification, these samples are stored. The de-identified samples are collected under various project-based schemes. The quality standards of our samples are established by two factors: pre-analytical quality control efforts, and record keeping of sample history by sample tracking system. All the samples have been phlebotomized under strict control of pre-analytical requirements. By record keeping of sample history with the sample tracking system, we can tell the exact temperature and elapsed time in various situations since the phlebotomy procedure or on arrival at the clinical laboratory. In this article, we will disclose how we have dealt with the de-identification procedure and sample quality.
Japanese Journal of Clinical Oncology 02/2011; 41(2):295-8. · 1.78 Impact Factor
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Yoshiki Kozu,
Koji Tsuta,
Takashi Kohno,
Ikuo Sekine,
Akihiko Yoshida,
Shunichi Watanabe,
Tomohide Tamura,
Jun Yokota,
Kenji Suzuki,
Hisao Asamura, Koh Furuta,
Hitoshi Tsuda
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ABSTRACT: Among the mutations of epidermal growth factor receptor (EGFR), deletions in exon 19 (DEL), and point mutations in exon 21 (L858R) predict the response to EGFR-tyrosine kinase inhibitors (TKIs) in primary lung adenocarcinoma. The ability to detecting such mutations using immunohistochemistry (IHC) would be advantageous.
The molecular-based and IHC-based EGFR mutations were analyzed in 577 lung adenocarcinomas using high resolution melting analysis (HRMA) and 2 mutation-specific antibodies, respectively.
In the molecular-based analyses, DEL was detected in 135 cases (23%), and L858R was detected in 172 cases (30%). In the IHC-based analyses, a positive reaction was detected in 59 cases (10%) for the DEL-specific antibody, and in 139 cases (24%) for the L858R-specific antibody. With the molecular-based results set as the gold standard, the sensitivity and specificity of the DEL-specific antibody were 42.2% and 99.5%, respectively, while the sensitivity and specificity of the L858R-specific antibody were 75.6% and 97.8%, respectively. The antibody specificities improved when the threshold for the mutation-positive reactions was set as >50% of immunopositive tumor cells. The significant predictors of the clinical response to EGFR-TKI were molecular-based EGFR mutations (p<0.001) and IHC-based EGFR mutations (p=0.001). However, a multivariate analysis revealed that only molecular-based EGFR mutations were significantly correlated with the clinical response (p<0.001).
Mutation-specific antibodies demonstrated extremely high specificities, but their sensitivities were not higher than those of molecular-based analyses. However, IHC should be performed before a molecular-based analysis, because it is more cost-effective and can effectively select candidates for EGFR-TKI therapy.
Lung cancer (Amsterdam, Netherlands) 12/2010; 73(1):45-50. · 3.14 Impact Factor
